Protocol : a method to study the direct reprogramming of lateral root primordia to fertile shoots

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Kareem , A , Radhakrishnan , D , Wang , X , Bagavathiappan , S , Trivedi , Z B , Sugimoto , K , Xu , J , Mähonen , A P & Prasad , K 2016 , ' Protocol : a method to study the direct reprogramming of lateral root primordia to fertile shoots ' , Plant Methods , vol. 12 , 27 . https://doi.org/10.1186/s13007-016-0127-5

Title: Protocol : a method to study the direct reprogramming of lateral root primordia to fertile shoots
Author: Kareem, Abdul; Radhakrishnan, Dhanya; Wang, Xin; Bagavathiappan, Subhikshaa; Trivedi, Zankhana B.; Sugimoto, Kaoru; Xu, Jian; Mähonen, Ari Pekka; Prasad, Kalika
Contributor: University of Helsinki, Biosciences
University of Helsinki, Institute of Biotechnology
Date: 2016-05-12
Language: eng
Number of pages: 14
Belongs to series: Plant Methods
ISSN: 1746-4811
URI: http://hdl.handle.net/10138/175911
Abstract: Background: Plants have the remarkable property to elaborate entire body plan from any tissue part. The conversion of lateral root primordium (LRP) to shoot is an ideal method for plant propagation and for plant researchers to understand the mechanism underlying trans-differentiation. Until now, however, a robust method that allows the efficient conversion of LRP to shoot is lacking. This has limited our ability to study the dynamic phases of reprogramming at cellular and molecular levels. Results: Here we present an efficient protocol for the direct conversion of LRP to a complete fertile shoot system. This protocol can be readily applied to the various ecotypes of Arabidopsis. We show that, the conversion process is highly responsive to developmental stages of LRP and changes in external environmental stimuli such as temperature. The entire conversion process can be adequately analyzed by histological and imaging techniques. As a demonstration, using a battery of cell fate specific markers, we show that confocal time-lapse imaging can be employed to uncover the early molecular events, intermediate developmental phases and relative abundance of stem cell regulators during the conversion of LRP to shoot. Conclusion: Our method is highly efficient, independent of genotypes tested and suitable to study the reprogramming of LRP to shoot in intact plants as well as in excised roots.
Subject: Arabidopsis
Auxin
Cytokinin
Lateral root
Shoot
Regeneration
Trans-differentiation
STEM-CELL NICHE
PLETHORA TRANSCRIPTION FACTORS
ARABIDOPSIS ROOT
TISSUE-CULTURE
MERISTEM
REGENERATION
INITIATION
THALIANA
WUSCHEL
MORPHOGENESIS
1183 Plant biology, microbiology, virology
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